ABSTRACT
We describe the separation of an active glutamate dehydrogenase [GDH (NADP+)] enzyme from the plasma of patients with P. falciparum infection using columns of sepharose anti-GDH (NADP+) of Proteus spp. The activity of this enzyme was also detected in P. falciparum culture supernatant. The parasitic origin of this enzyme was suggested by western blot analysis using anti-P. falciparum culture supernatant and anti-whole parasite antibodies. The differential inhibition of the P. falciparum GDH (NADP+) indicates that some epitopes recognised by the antibodies in both preparations may be different. The determination of P. falciparum GDH (NADP+) activity could be developed into a specific technique for the diagnosis of falciparum malaria.
Subject(s)
Animals , Glutamate Dehydrogenase/blood , Humans , Malaria, Falciparum/blood , Plasmodium falciparum/enzymologyABSTRACT
The major aim of this study was to characterize a soluble Plasmodium falciparum antigen from the plasma of malaria-infected humans and Plasmodium falciparum culture supernatants, using immunoabsorbent techniques and Western blotting. An Mr 60-kDa protein was isolated from the plasma of patients with Plasmodium falciparum malaria by affinity chromatography using rabbit anti-Proteus spp GDH(NADP+) serum as ligand. This protein, present in plasma of patients with acute Plasmodium falciparum infection, in Plasmodium falciparum culture supernatants, and in immune complexes, was tested with Plasmodium falciparum malaria hyperimmune serum from patients living in hyperendemic areas and rabbit anti-Proteus spp GDH(NADP+) serum prepared in the laboratory. In this report, we describe the results of a study showing that parasite GDH(NADP+) can be used to detect the presence of Plasmodium falciparum. It appears that this technique permits the chromatographic detection of a Plasmodium falciparum excretion antigen that may be used in the production of monoclonal antibodies to improve immunodiagnostic assays for the detection of antigenemia, and opens the possibility of its use as a non-microscopic screening method.
Subject(s)
Humans , Animals , Rabbits , Antigens, Protozoan/blood , Glutamate Dehydrogenase/blood , Malaria, Falciparum/enzymology , Plasmodium falciparum/enzymology , Acute Disease , Antibodies, Bacterial/blood , Antibodies, Monoclonal , Blotting, Western , Chromatography, Affinity , Culture Media , Glutamate Dehydrogenase/immunology , Glutamate Dehydrogenase/isolation & purification , Immunoglobulin G/blood , Immunosorbent Techniques , Malaria, Falciparum/blood , Malaria, Falciparum/parasitology , Plasmodium falciparum/isolation & purification , SolubilityABSTRACT
In this study, glutamate dehydrogenase [GLDH] activity was evaluated spectrophotometrically in 100 individuals divided into four groups: A normal group [GN], diabetic group [GD], diabetic group accompanied with renal failure and diabetic group accompanied with myocardial infarction [GDM]. Serum aspartate aminotransferase [AST] activity in normal group was 27.9 U/L; while in case of GDM group, it increased significantly to be 125.64 U/L. Serum alanine aminotransferase [ALT] activity in the normal group was 28.76 U/L and it was slightly changed in the other diabetic groups
Subject(s)
Humans , Male , Female , Diabetes Mellitus/complications , Glutamate Dehydrogenase/blood , Aspartate Aminotransferases/blood , Alanine Transaminase/bloodSubject(s)
Humans , Male , Female , Schistosomiasis/blood , Glutamate Dehydrogenase/blood , /blood , Liver Function Tests/methodsABSTRACT
The serum activity of glutamate dehydrogenase [GLDH] was measured spectrophotometrically in 75 cases, normal healthy 25 cases as a control group [I], chronic renal failure 25 cases group [II] and renal transplantation patients 25 cases group [III]. The mean level of GLDH activity in chronic renal failure [12.55 U/L] was significantly higher than the control group [1.58 U/L], and also, higher than the corresponding mean level of renal transplantation patients [2.54 U/L]. The result indicated that evaluation of serum GLDH activity has diagnostic potential